Targeted gene correction minimally impacts whole-genome mutational load in human-disease-specific induced pluripotent stem cell clones

Keiichiro Suzuki, Chang Yu, Jing Qu, Mo Li, Xiaotian Yao, Tingting Yuan, April Goebl, Senwei Tang, Ruotong Ren, Emi Aizawa, Fan Zhang, Xiuling Xu, Rupa Devi Soligalla, Feng Chen, Jessica Kim, Na Young Kim, Hsin Kai Liao, Chris Benner, Concepcion Rodriguez Esteban, Yabin JinGuang Hui Liu*, Yingrui Li, Juan Carlos Izpisua Belmonte

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

133 Scopus citations


The utility of genome editing technologies for disease modeling and developing cellular therapies has been extensively documented, but the impact of these technologies on mutational load at the whole-genome level remains unclear. We performed whole-genome sequencing to evaluate the mutational load at single-base resolution in individual gene-corrected human induced pluripotent stem cell (hiPSC) clones in three different disease models. In single-cell clones, gene correction by helper-dependent adenoviral vector (HDAdV) or Transcription Activator-Like Effector Nuclease (TALEN) exhibited few off-target effects and a low level of sequence variation, comparable to that accumulated in routine hiPSC culture. The sequence variants were randomly distributed and unique to individual clones. We also combined both technologies and developed a TALEN-HDAdV hybrid vector, which significantly increased gene-correction efficiency in hiPSCs. Therefore, with careful monitoring via whole-genome sequencing it is possible to apply genome editing to human pluripotent cells with minimal impact on genomic mutational load.

Original languageEnglish (US)
Pages (from-to)31-36
Number of pages6
JournalCell Stem Cell
Issue number1
StatePublished - Jul 3 2014
Externally publishedYes

Bibliographical note

Funding Information:
We would like to thank K. Mitani, P. Ng, and A. Lieber for HDAdV production; L. de Oñate for technical help; and M. Schwarz, P. Schwarz, and R. Chen for administrative help. G.H.L., J.Q., and X.X. were supported by National Basic Research Program (973 program: 2014CB910500, 2014CB964600), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA01020312), NSFC (81271266, 31222039, 81330008, 81371342, 31201111, 81300261, and 81300677), Key Research Program of the Chinese Academy of Sciences (KJZD-EW-TZ-L05), Beijing Natural Science Foundation (7141005 and 5142016), the Thousand Young Talents program of China, National Laboratory of Biomacromolecules (2013kf05, 2013kf1, and 2014kf02), and State Key Laboratory of Drug Research (SIMM1302KF-17). K.S. and M.L. were supported by a California Institute for Regenerative Medicine training fellowship. Work in the laboratory of J.C.I.B. was supported by the G. Harold and Leila Y. Mathers Charitable Foundation, The Leona M. and Harry B. Helmsley Charitable Trust (#2012-PG-MED002), The Glenn Foundation, CIRM, and NIH (5U01HL107442).

ASJC Scopus subject areas

  • Molecular Medicine
  • Genetics
  • Cell Biology


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